Epithelial Cancer is an insidious disease. For instance, one type of epithelial cancer, carcinoma of the bladder, represents a significant source of morbidity and mortality. Bladder cancer reportedly ranks 10th in males and 12th in females in cancer related mortality. Therapies available for the treatment of bladder cancer include adjuvant chemotherapy or immunotherapy, transurethral resection of superficial disease, radical cystectomy or radiotherapy which is often combined with systemic chemotherapy. Despite these therapeutic options, overall survival has not changed appreciably. Thus, new therapeutic modalities must be developed for the treatment of bladder cancer.
Gene therapy strategies have been reportedly developed as an alternative therapeutic approach. Distinct approaches have been developed to treat neoplasms based on gene transfer methods. Methods have been reportedly developed to correct specific lesions at defined genetic loci which give rise to neoplastic transformation and progression. Overexpression of dominant oncogenes can be addressed using techniques to inhibit the transforming gene or gene product. It has been reported that loss of tumor suppressor gene function may be approached using methods to reconstitute wild-type tumor suppressor gene. Besides these methods to achieve mutation compensation, genetic techniques have been reportedly developed to specifically and selectively eradicate tumor cells. These approaches of molecular chemotherapy reportedly rely on specific expression of toxin genes in neoplastic cells. Finally, gene transfer methods have been reportedly used to achieve antitumor immunization. These methods of genetic immunopotentiation reportedly use techniques of genetic immunoregulation to enhance immune recognition of tumors. Consequently, a variety of distinct approaches reportedly have been developed to accomplish gene therapy of cancer.
A high incidence of mutations has reportedly been observed in tumor suppressor genes, such as p53 and RB, in the case of carcinoma of the bladder. For such genetic lesions of tumor suppressor genes, reversion of the neoplastic phenotype can be demonstrated with replacement of the corresponding wild-type tumor suppressor gene.
In vitro studies using cell lines derived from human bladder tissues have reportedly demonstrated efficient transgene expression following infection with recombinant adenovirus. Experiments in vivo reportedly have also shown adenovirus transgene expression in the urinary bladder of rodents after intravesical delivery. In vitro experiments with wild-type adenovirus demonstrate that virus attachment and internalization is not influenced by benzyl alcohol, but do reportedly demonstrate an enhanced uncoating of the virion. In vivo efforts with agents (e.g. acetone, DMSO, prolamine sulfate) can reportedly break down the protective “mucin” layer that protects the bladder epithelium from bacteria, viruses and other pathogens.
U.S. Pat. No. 5,789,244 claims a composition comprising a viral vector in which a nucleotide sequence encoding a transgene has been inserted, wherein the viral vector is formulated in a buffer comprising ethanol in a concentration range of about 1% to 50% (v/v). U.S. Pat. No. 5,837,520 claims a method for purification of an intact viral particle from a cell lysate comprising treating the cell lysate which contains the intact viral particle with an enzymatic agent that selectively degrades both unencapsulated DNA and RNA; chromatographing the treated lysate from the first step on a first resin; and chromatographing the eluant from the second step on a second resin; wherein one resin is an anion exchange resin and the other is an immobilized metal ion affinity resin. U.S. Pat. No. 5,932,210 describes a composition comprising a recombinant adenovirus expression vector and a pharmaceutically acceptable carrier, the vector comprising: (a) an insert of exogenous DNA comprising a gene encoding a foreign protein; and (b) adenovirus DNA in which all of the coding sequences of E1a, E1b, and protein IX, and at least part of E3 have been deleted. U.S. Pat. No. 6,165,779 discloses a composition comprising a recombinant virus vector formulated in a buffer comprising a detergent. U.S. Pat. No. 6,210,939 claims a recombinant adenovirus expression vector comprising (a) an insert of exogenous DNA comprising a gene encoding a foreign protein and (b) adenovirus DNA which has sustained a deletion beginning at nucleotide 357 and ending at nucleotide 4020 to 4050. Finally, U.S. Pat. No. 6,312,681 discloses a method for delivering an adenoviral vector which comprises a transgene to a cancer cell in the epithelial membrane of a bladder, the method comprising administering to the epithelial membrane the adenoviral vector and between 1% and 50% (v/v) ethanol, wherein the adenoviral vector infects the cell and the transgene is expressed in infected cells. All of these references are hereby incorporated by reference thereto in their entirety.
Notwithstanding the foregoing, there exists a need for formulations for therapeutic use that improve the efficiency of the transgene delivery. Vectors that are unstable present difficulty in administering the desired therapeutic agent to the patient. Because of in vivo instability, there is a need for vector stabilization such that there is an increase in the transduction of the therapeutic agent that is to be administered.